CHARACTERIZATION OF THE GENE FOR AN IMMUNODOMINANT 72 KDA LIPOPROTEINOF MYCOPLASMA-MYCOIDES SUBSP MYCOIDES SMALL COLONY TYPE

With the aim of characterizing specific immunogenic proteins of Mycopl asma mycoides subsp, mycoides small colony (SC) type, the aetiological agent of contagious bovine pleuropneumonia, a gene encoding a major i mmunogenic protein of 72 kDa named P72 was cloned and expressed in Esc herichia coli. The expressed protein was of the same apparent molecula r mass as that produced by the parent strain. The predicted molecular mass of P72, based on the DNA-deduced amino acid sequence. was 61.118 kDa, significantly lower than the apparent molecular mass of endogenou s or recombinant P72 on SDS-PAGE. Analysis of the amino acid sequence revealed a typical prokaryotic signal peptidase II - membrane lipoprot ein lipid attachment site and a transmembrane structure domain in the leader sequence at the aminoterminal end of the protein. P72 was shown to be a lipoprotein and its surface location was confirmed by trypsin treatment of whole cells. An unassigned gene encoding a peptide with some similarity to P72 was found on the genome sequence of M. capricol um subsp. capricolum but not on that of Mycoplasma genitalium. The P72 gene was detected in 11/11 M. mycoides subsp. mycoides SC strains. An tiserum against recombinant P72 reacted strongly with 12/12 strains of M. mycoides subsp. mycoides SC, weakly with Mycoplasma bovine group 7 strain PG50, but not with other members of the 'mycoides cluster' or closely related mycoplasmas. Cows experimentally contact-infected with M. mycoides subsp. mycoides SC developed a humoral response against P 72 within 35 d. P72 is a specific antigenic membrane lipoprotein of M. mycoides subsp. mycoides SC with potential for use in development of diagnostic reagents. It seems to belong to a family of lipoproteins of the 'mycoides cluster'.